Involvement of RBP-J interacting and tubulin-associated protein in the distribution of protein regulator of cytokinesis 1 in mitotic spindles
The protein regulator of cytokinesis 1 (PRC1) is a key regulator of microtubule crosslinking and bundling, which is crucial for spindle formation and cytokinesis. RITA, the RBP-J interacting and tubulin-associated protein, is a microtubule associated protein. We have reported that RITA localizes to...
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2024.1472340/full |
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| author | Julia Caspers Andreas Ritter Badi Bahrami Samira Catharina Hoock Susanne Roth Alexandra Friemel Franz Oswald Frank Louwen Nina-Naomi Kreis Juping Yuan |
| author_facet | Julia Caspers Andreas Ritter Badi Bahrami Samira Catharina Hoock Susanne Roth Alexandra Friemel Franz Oswald Frank Louwen Nina-Naomi Kreis Juping Yuan |
| author_sort | Julia Caspers |
| collection | DOAJ |
| description | The protein regulator of cytokinesis 1 (PRC1) is a key regulator of microtubule crosslinking and bundling, which is crucial for spindle formation and cytokinesis. RITA, the RBP-J interacting and tubulin-associated protein, is a microtubule associated protein. We have reported that RITA localizes to mitotic spindles modulating microtubule dynamics and stability as well as to spindle poles affecting the activity of Aurora A. As defective chromosome congression and segregation are the most remarkable features of cells depleted of RITA, we aimed to explore further potential related mechanisms, using various cellular and molecular techniques, including clustered regularly interspaced short palindromic repeats technique/deactivated CRISPR-associated protein 9 (CRISPR/dCas9), mass spectrometry, confocal microscopy, immunofluorescence, immunoprecipitation and Western blot analysis. Here, we show that FLAG-RITA precipitates PRC1 and tubulin, and that these two proteins co-localize in the central region of the central spindle. Reduction of RITA enlarges the staining area of PRC1 in mitotic spindles as well as in the central spindle. Its suppression reduces the inter-centromere distance in metaphase cells. Interestingly, microtubule bundles of the central spindle are often less organized in a non-parallel pattern, as evidenced by increased angles, relative to corresponding separating chromosomes. These data suggest a novel role for RITA in mitotic distribution of PRC1 and that its deregulation may contribute to defective chromosome movement during mitosis. As both RITA and PRC1 are closely associated with malignant progression, further work is required to elucidate the detailed molecular mechanisms by which RITA acts as a modulator in central spindle formation and cytokinesis. |
| format | Article |
| id | doaj-art-e9ff24aac7c1403ab01514e62489d010 |
| institution | Kabale University |
| issn | 2296-634X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj-art-e9ff24aac7c1403ab01514e62489d0102025-01-07T06:49:06ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2025-01-011210.3389/fcell.2024.14723401472340Involvement of RBP-J interacting and tubulin-associated protein in the distribution of protein regulator of cytokinesis 1 in mitotic spindlesJulia Caspers0Andreas Ritter1Badi Bahrami2Samira Catharina Hoock3Susanne Roth4Alexandra Friemel5Franz Oswald6Frank Louwen7Nina-Naomi Kreis8Juping Yuan9Obstetrics and Prenatal Medicine, Department of Gynecology and Obstetrics, University Hospital Frankfurt, J. W. Goethe-University, Frankfurt, GermanyObstetrics and Prenatal Medicine, Department of Gynecology and Obstetrics, University Hospital Frankfurt, J. W. Goethe-University, Frankfurt, GermanyObstetrics and Prenatal Medicine, Department of Gynecology and Obstetrics, University Hospital Frankfurt, J. W. Goethe-University, Frankfurt, GermanyObstetrics and Prenatal Medicine, Department of Gynecology and Obstetrics, University Hospital Frankfurt, J. W. Goethe-University, Frankfurt, GermanyObstetrics and Prenatal Medicine, Department of Gynecology and Obstetrics, University Hospital Frankfurt, J. W. Goethe-University, Frankfurt, GermanyObstetrics and Prenatal Medicine, Department of Gynecology and Obstetrics, University Hospital Frankfurt, J. W. Goethe-University, Frankfurt, GermanyCenter for Internal Medicine, Department of Internal Medicine I, University Medical Center Ulm, Ulm, GermanyObstetrics and Prenatal Medicine, Department of Gynecology and Obstetrics, University Hospital Frankfurt, J. W. Goethe-University, Frankfurt, GermanyObstetrics and Prenatal Medicine, Department of Gynecology and Obstetrics, University Hospital Frankfurt, J. W. Goethe-University, Frankfurt, GermanyObstetrics and Prenatal Medicine, Department of Gynecology and Obstetrics, University Hospital Frankfurt, J. W. Goethe-University, Frankfurt, GermanyThe protein regulator of cytokinesis 1 (PRC1) is a key regulator of microtubule crosslinking and bundling, which is crucial for spindle formation and cytokinesis. RITA, the RBP-J interacting and tubulin-associated protein, is a microtubule associated protein. We have reported that RITA localizes to mitotic spindles modulating microtubule dynamics and stability as well as to spindle poles affecting the activity of Aurora A. As defective chromosome congression and segregation are the most remarkable features of cells depleted of RITA, we aimed to explore further potential related mechanisms, using various cellular and molecular techniques, including clustered regularly interspaced short palindromic repeats technique/deactivated CRISPR-associated protein 9 (CRISPR/dCas9), mass spectrometry, confocal microscopy, immunofluorescence, immunoprecipitation and Western blot analysis. Here, we show that FLAG-RITA precipitates PRC1 and tubulin, and that these two proteins co-localize in the central region of the central spindle. Reduction of RITA enlarges the staining area of PRC1 in mitotic spindles as well as in the central spindle. Its suppression reduces the inter-centromere distance in metaphase cells. Interestingly, microtubule bundles of the central spindle are often less organized in a non-parallel pattern, as evidenced by increased angles, relative to corresponding separating chromosomes. These data suggest a novel role for RITA in mitotic distribution of PRC1 and that its deregulation may contribute to defective chromosome movement during mitosis. As both RITA and PRC1 are closely associated with malignant progression, further work is required to elucidate the detailed molecular mechanisms by which RITA acts as a modulator in central spindle formation and cytokinesis.https://www.frontiersin.org/articles/10.3389/fcell.2024.1472340/fullRITAmitotic defectscytokinesis failurePRC1the central spindle |
| spellingShingle | Julia Caspers Andreas Ritter Badi Bahrami Samira Catharina Hoock Susanne Roth Alexandra Friemel Franz Oswald Frank Louwen Nina-Naomi Kreis Juping Yuan Involvement of RBP-J interacting and tubulin-associated protein in the distribution of protein regulator of cytokinesis 1 in mitotic spindles Frontiers in Cell and Developmental Biology RITA mitotic defects cytokinesis failure PRC1 the central spindle |
| title | Involvement of RBP-J interacting and tubulin-associated protein in the distribution of protein regulator of cytokinesis 1 in mitotic spindles |
| title_full | Involvement of RBP-J interacting and tubulin-associated protein in the distribution of protein regulator of cytokinesis 1 in mitotic spindles |
| title_fullStr | Involvement of RBP-J interacting and tubulin-associated protein in the distribution of protein regulator of cytokinesis 1 in mitotic spindles |
| title_full_unstemmed | Involvement of RBP-J interacting and tubulin-associated protein in the distribution of protein regulator of cytokinesis 1 in mitotic spindles |
| title_short | Involvement of RBP-J interacting and tubulin-associated protein in the distribution of protein regulator of cytokinesis 1 in mitotic spindles |
| title_sort | involvement of rbp j interacting and tubulin associated protein in the distribution of protein regulator of cytokinesis 1 in mitotic spindles |
| topic | RITA mitotic defects cytokinesis failure PRC1 the central spindle |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2024.1472340/full |
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